title: Determination of malondialdehyde in biological fluids by high-performance liquid chromatography using rhodamine B hydrazide as the derivatization reagent.
abstract: Malondialdehyde (MDA) is a biomarker for lipid peroxidation, and studies of sensitive and selective analytical methods for it are very important for pathological research. The aim of this work was to develop and validate a novel HPLC method for the quantification of MDA in biological fluids using rhodamine B hydrazide (RBH) as the derivatization reagent. After pretreatment and derivatization in acid medium at 50 °C for 40 min, the RBH-derivatized MDA was separated on a Kromasil C18 column at 25 °C and detected by a fluorescence detector at excitation wavelength of 560 nm and emission wavelength of 580 nm. The results showed linearity in the range of 0.8-1500.0 nM with a detection limit of 0.25 nM (S/N = 3). The recovery of MDA from plasma and urine was 91.50 to 99.20%, with a relative standard deviation range of 1.45 to 3.26%. In comparison to other methods reported for the determination of MDA, the proposed method showed superiority in simplicity, more sensitivity, shorter derivatization time, and less interference. The developed method was applied to quantification of MDA in human biological fluids collected from five volunteers with a concentration range of 24.62-245.00 nM.
authors.affiliation: College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Pingliang
authors.initials: P
authors.lastname: Li
authors.affiliation: College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Guanglong
authors.initials: G
authors.lastname: Ding
authors.affiliation: College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Yufang
authors.initials: Y
authors.lastname: Deng
authors.affiliation: College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Darunee
authors.initials: D
authors.lastname: Punyapitak
authors.affiliation: College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Deguang
authors.initials: D
authors.lastname: Li
authors.affiliation: College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, China. Electronic address: caoysong@126.com.
authors.forename: Yongsong
authors.initials: Y
authors.lastname: Cao
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D006834:Hydrazines; D008297:Male; D008315:Malondialdehyde / Q000032:analysis*; D012235:Rhodamines
chemical_list: D006834:Hydrazines; D012235:Rhodamines; C543681:rhodamine B hydrazide; D008315:Malondialdehyde
keywords: Determination; Free radicals; High-performance liquid chromatography; Malondialdehyde; Precolumn derivatization; Rhodamine B hydrazide